The 11α-hydroxylation of 16α, 17-epoxyprogesterone (EP) catalyzed by Rhizopus nigricans is crucial for the steroid industry. However, lower conversion rate of the biohydroxylation restricts its potential industrial application. The 11α-steroid hydroxylase CYP509C12 from R. oryzae were reported to play a crucial role in the 11α-hydroxylation in recombinant fission yeast. In the present study, the CYP509C12 of R. oryzae (RoCYP) was introduced into R. nigricans using the liposome-mediated mycelial transformation. Heterologous expression of RoCYP resulted in increased fungal growth and improved intracellular reactive oxygen species content in R. nigricans. The H₂O₂ levels in RoCYP transformants were approximately 2-folder that of the R. nigricans wild type (RnWT) strain, with the superoxide dismutase activities increased approximately 45% and catalase activities decreased approximately 68%. Furthermore, the 11α-hydroxylation rates of EP in RoCYP transformants (C4, C6 and C9) were 39.7%, 38.3% and 38.7%, which were 12.1%, 8.2% and 9.4% higher than the rate of the RnWT strain, respectively. This paper investigated the effect of heterologous expression of RoCYP in R. nigricans, providing an effective genetic method to construct the engineered strains for steroid industry.

黑根霉菌催化的16α，17-环氧孕酮（EP）的11α-羟基化对于类固醇工业至关重要。然而，较低的生物羟基化转化率限制了其潜在的工业应用。据报道，米曲霉的11α-甾类羟化酶CYP509C12在重组裂变酵母中的11α-羟化中起关键作用。在本研究中，CYP509C12的米根霉（RoCYP）引入ř。黑体使用脂质体介导的菌丝体转化。RoCYP的异源表达导致真菌的生长和R中细胞内活性氧含量的增加。黑农人。为H _2个ö ₂在水平RoCYP转化体用大约2文件夹，所述的R.皮症野生型（RnWT）株，与超氧化物歧化酶活性增加约45％的过氧化氢酶和活性下降约68％。此外，RoCYP转化子（C4，C6和C9）中EP的11α-羟基化率分别为39.7％，38.3％和38.7％，分别比RnWT菌株高12.1％，8.2％和9.4％。本文研究了RoCYP异源表达在小鼠中的作用R. nigricans，提供了有效的遗传方法来构建用于类固醇工业的工程菌株。